Comparative Cytotoxicity of Primary Rat Cardiomyocyte, Human Lung Fibroblast (W138) and Rabbit Corneal Epithelial (SIRC) Cell Cultures Exposed to Chloroform and Allyl Alcohol
Mastenbrook, M. Scott
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The purpose of this study was to determine the in vitro cytotoxic effects of chloroform and allyl alcohol in primary rat cardiomyocytes, human lung fibroblasts (WI38) and rabbit corneal epithelial (SIRC) cells. The cells were exposed to varying concentrations of chloroform or allyl alcohol (range 1.22 x 10 -1 to 6.0 x 10 -3 M) for 4 hrs. Cytotoxicity was assessed by morphological (phase contrast microscopy), functional (beating rates for cardiomyocytes; cell proliferation for WI38 and SIRC cells), and enzymatic criteria (CPK, LDH) at 4, 8, 24 or 48 hrs post-exposure. Chloroform was irreversibly toxic to all of the cell types at 4.9 x 10 -2 M, while a concentration of 2.4 x 10 -2 M produced milder toxic effects (i.e. loss of cardiomyocyte contraction and rhythmicity, slightly elevated LDH leakage); leaving the cells viable. Allyl alcohol, at 1.22 x 10 -1 M produced toxic effects relating to cell viability, cellular membrane injury, and proliferation; however, there was a slight recovery of the cells post-exposure to the xenobiotic. Cellular enzyme leakage assessment showed membrane damage at 6.0 x 10 -3 M. In this study, chloroform exposure caused acute cellular damage during the exposure period, while allyl alcohol demonstrated acute as well as delayed cytotoxicity in the cell systems investigated. Dose response curves revealed that a statistically significant relationship existed between dose and a response to that dose for the various cell types. This preliminary data suggests that appropriate cell culture systems provide a sensitive method for in vitro cytotoxicity assessment.